Standard reference reagent package



United States Patent O 3,549,297 STANDARD REFERENCE REAGENT PACKAGERoland J. Starkey, Jr., North Canton, Ohio, assignor to Ben VenueLaboratories, Inc., Pittsburgh, Pa., a corporation of Pennsylvania NoDrawing. Original application Mar. 31, 1966, Ser. No. 538,973, nowPatent No. 3,449,885, dated June 17, 1969. Divided and this applicationSept. 9, 1968, Ser. No. 758,627

The portion of the term of the patent subsequent to June 18, 1986, hasbeen disclaimed Int. Cl. B65d 85/00 U.S. Cl. 20646 9 Claims ABSTRACT OFTHE DISCLOSURE Method of providing a plurality of vials or packages ofviral antisera or reagent from a common lot, each of which has auniform, homogenous equal content and moisture content and a long shelflife, in lasting compliance with and adherence to the required standardand specifications, by treating the batch or lot of viral antiserum orreagent to be packaged or vialed in bulk by freeze drying (lyophilize)the same, if necessary, pulverizinig or grinding the dried serum to auniform size and consistency, vacuum drying the bulk lot of powderedserum, with stirring or agitation, if desired, to obtain a lot of serumhaving a uniform, homogenous, moisture content, powder filling theseparate vials or packages of the lot under sterile, moisture freeconditions to a uniform fill and content, and sealing the separate vialsor packages with a dry moisture free and proof seal.

CROSS-REFERENCE TO RELATED APPLICATION This application is a division ofapplicants copending application Ser. No. 538,973 filed Mar. 31, 1966,now US. Pat. No. 3,49,885.

SUMMARY AND DESCRIPTION OF THE INVENTION This invention relates to a newand improved method and process for packaging or vialing moisture freeviral antisera and other diagostic reagents, vaccines, and the like; toa new and improved method and process for packaging or vialing aplurality of containers, of uniform, homogeneous viral antisera andother diagnostic reagents, vaccines, and the like, from a single lotand, more particularly, to a new and improved method and process forpackaging or vialing viral antisera for use as research referencereagents and the like.

It is customary to package viral antisera, and the like, in vials fordistribution and use as standard reference reagents. Such vialedantisera is prepared to uniform standards in a plurality of vials orseparate packages from a given lot, batch or bulk supply with theexpectations that the antisera will remain up to standard in quantity,uniformity, quality and moisture content over extended periods of timesor for a long shelf life so that subsequent researchers and users areassured of reference, or other diagnostic, reagents for use in researchprojects, and the like, which, in fact, are sterile and complyaccurately with the standards established for the particular referencereagent as published and distributed to users of the reagent. One of themost important criteria which must be met upon original packaging orvialing of the antisera, or other reagnet, and which must be maintainedthroughout the shelf life of the respective vials of anti sera orreagent is moisture or, more appropriately, lack of moisture content.

Further, if the respective vials are to be true standards, in fact, themoisture content must be the same in each vial of a given lot and, ifthe reagents are to retain their standard characteristics throughouttheir anticipated shelf life, the moisture content of each vial must, infact, be below the maximum allowed from the time of sealing to the timeof use. Also, it is preferable that the vials of reagent not be reduced,during the drying process, to a moisture content below the maximumpermitted by the standard or specifications and then be permitted toincrease to a higher moisture content, even though the latter, highercontent be within the maximum allowed under the applicable standard andspecifications. The long accepted prior practice and custom in the art,for vialing viral antiserum is set forth, for example, in GeneralSpecifications for Lyophilization and Packaging of Research Reagents, asrevised Feb. 24, 1966, by the Research Reference Reagents Branch,National Institute of Allergy and Infectious Diseases, NatitonalInstitutes of Health.

Analyses and studies of previously packaged and vialed standardreference viral antisera have established that vials of viral antisera,which have been prepared by freeze drying and sealing under vacuum andaccumulated in the expectation that they would and could function and,in fact, be standard reference reagents having a long shelf life suchthat the standard and specifications of the respective vials would existand continue to exist over long periods of time, while under acceptableand accepted storage and shelf life conditions, frequently had amoisture content exceeding specifications or standard and that theindividual vials varied considerably in dried sera content, apparentlybecause of differences in original moisture content. And, this has beenso even though the samples, which were tested during the originalpreparation of the vials, were Within the prescribed standard andspecifications.

The general object of this invention is, therefore, to provide a new andimproved method and process for packaging or vialing viral antisera andother diagnostic reagents, vaccines, and the like, for use as standardreference reagents, and other purposes, wherein individual packages of agiven lot are uniform in moisture, quantity and quality content.

Another object of this invention is to provide a new and improved methodand process for packaging or vialing viral antisera, and the like,within the meaning of this invention, which ensures that the content andcharacteristics of each vial of each lot of antisera or reagent is thesame or substantially the same and has a homogenous residual moisturecontent which is within and remains within the permissiblespecifications and standard throughout the life of the vial.

Still other objects of this invention include the provision of a new andimproved method and process for packaging or vialing viral antisera, andthe like, within the meaning of this invention, for use as standardreference reagents, and other appropriate purposes, which is efiicientand effective; which ensures that the vials or packages of apredetermined lot have a full, properly moisture free content; whichprovides even, effective and proper drying of the entire lot to a commonhomogenous residual moisture content so that the reagent in one vial orpackage does not differ in a meaningful way in moisture content fromthat in the other vials or packages of the same lot; which provides animproved and proper drying procedure; which prevents the reintroductionof moisture into the contents of the respective vials or packages, afterpackaging is complete; which ensures a properly dried and sterilefilling, to constant fill level, of each vial or package of a given lot;and, which 3 provides vialed or packaged vial antisera or otherreference reagents, and the like, which are the same for each lot, andwhich have a long shelf life and maintain their standard andspecifications throughoutsuch shelf life.

Still furtherobjects of this invention include the provision of a newand improved method and process for packaging or vialing 'viral antiserastandard reference reagents, and the like, within the meaning of thisinvention which is adapted for use with standard or substantiallystandard or easily modified apparatus; which is safe and foolproof inoperation; which does not contaminate or otherwise adversely affect thereagent; and, which is useful for lots ofreagent of a wide initial rangeof moisture characteristics and volume.

A still further object of this invention is to provide a. new andimproved method and process for packaging or vialing viral antisera, andthe like, Within the meaning of this invention, obtaining one or more ofthe objects set forth above.

These and other objects and advantages of this invention will appearfrom the following description of preferred and modified forms thereof.

Briefly the process of this invention, in contrast with prior art, andpractice followed with respect to the packaging or vialing of viralantisera standard reference reagents, and the like, within the meaningof this invention, comprises providing a plurality of vials or packagesof viral antisera or reagent from a common lot, each of which has auniform, homogenous equal content and moisture content and a long shelflife, in lasting compliance with and adherence to the required standardand specifications, by treating the batch or lot of viral antiserum orreagent tobe packaged or vialed in bulk, to freeze dry (lyophilize) thesame; pulverizing or grinding the freeze dried serum to a uniform sizeand consistency; vacuum drying the bulk lot of powdered serum, withstirring or agitation, if desired, to obtain a lot of serum having auniform, homogenous, moisture content; powder filling the separate vialsor packages of the lot under sterile, moisture free conditions to auniform fill and content; and sealing the separate vials or packageswith a dry moisture free, and proof seal.

In those instances where the moisture content of the lot or batch ofserum or reagent is initially sufiiciently low, the lot may bepulverized or ground before lyophilizing or, if the moisture content issufiiciently low, the lyophilizing step may be dispensed with, providedthe objects and purposes of this invention are otherwise obtained.

' The method and process of this invention is adapted for use eitherwith a starting serum, diagnostic reagent, vaccine, and the like, whichis a frozen (or unfrozen, for that matter) liquid or solution, or asolid of a unitary,

.monolithic or divided granular or larger nature, but, not

suitable for packaging or rvialing because, interalia, of a moisturecontent exceeding specifications or standard.

t The method and process of this invention while particularly adaptedfor packagaing or vialing viral antisera standard reference reagents mayhave a wide range of applications in the preparation, from a single lotof a' reagent, within the meaning of this invention, having excessivemoisture content of a plurality of separate packages, vials, ampoules,or the like, each of which has a uniform content of reagent having anhomogenous moisture content and a long shelf life, during which themoisture content remains stable and the reagent does not depart from theset and expected specifications and standard.

The invention is exemplified below by the detailing of a preferred formthereof particularly adapted for use in preparing vialed viral antiserafor use as standard reference reagents and by pointing out modificationsto the preferred form of this invention by which bacterial antisera,vaccines and other reagents, within the meaning of the invention, mayalso be provided as a plurality of vialed i or packaged units containinguniform, homogenous moisture free contents from a single lot.

Viral antisera for standard reference reagents is generally provided asa frozen solution in a bulk package which is thawed or otherwisereleased from the package, in which it is shipped, and filtered, bothunder sterile conditions.

In accordance with the precepts of this invention the batch or lot offiltered viral antisera is freeze dried or lyophilized, to eliminate thewater of solution and reduce the moisture content sufliciently to permitthe carrying out and accomplishment of the other steps of the method andprocess of this invention. Lyophilizing is accomplished generally in amanner known to the art and specifically for 'viral antiserum may beaccomplished as set forth in the Bulletin of the Parenteral DrugAssociations, vol. 20, No. 2, March-April 1966, pp. 39-47. I

After lyophilizing is completed, jthe bulk antiserum is ground orcomminuted, under sterile conditions, into a fine homogenous powder,preferably of about mesh.

The powdered antiserum is then vacuum dried, in sterile apparatus, overa suitable desiccant, preferably molecular sieve, until, upon testing,the moisture content of the bulk antiserum has been reduced to thatrequired by the standard or specifications.

Drying of the antiserum in bulk and in powdered form to the desiredstandard and specifications provide a homogenous serum lot which is muchmore quickly and evenly dried and freed of its moisture content, becauseof the homogenous physical nature of the antiserum when in powderedform, and the large surface and of the powder vis-a-vis the totalquantity of antiserum in the lot.

For viral antiserum, the vacuum chamber is preferably flushed withnitrogen before use. The viral antiserum lot is dried in vacuo at 25 C.for about thirty-six hours or until the moisture content at the cornersand center (and at and below the surface) of the tray or bulk containerof powdered antiserum being dried not only meets specifications but isthe same within plus or minus 0.2%. Note, when the volume of powderedantiserum is such that a plurality of trays or containers of powderedantiserum are prepared for vacuum drying in a single drier, that thesamples from all of the trays must assay within plus or minus 0.2% inorder to ensure that the antiserum is homogenous in moisture content.Further, the contents of the separate trays are intermixed: beforefilling of the separate vials or packages in accordance with theprecepts of this invention, as explained below. 7

Also, if desired, the powdered antiserum may be stirred or agitatedduring vacuum drying to increase the drying rate and accelerate andensure obtaining of homogenous conditions throughout the powder mass.

After vacuum drying, the antiserum of a given lot or batch is filledinto a plurality of separate vials or packages. Pending filling, thedried antiserum may be stored, if desired, in sterile, dry stainlesssteel beakers in a sterile, anhydrous atmosphere in a desiccator overmolecular sieve.

The antiserum of a given lot is filled, under sterile conditions, in aconventional, for example, preferably high speed, filling apparatus.Each vial, for example, is first thoroughly dried and then purged withsterile anhydrous nitrogen, or other suitable, inert gas immediatelybefore filling. Further, the vials are blanketed with the same or asimilar sterile, anhydrous inert gas until sealed and a positivepressure and protective atmosphere is maintained on and about the powderin the filling machine with the same or a similar sterile, anhydrousinert gas. Each vial is filled to the predetermined amount and, sincethe powder is homogenous and the antiserum reduced to prescribedmoisture content according to a predetermined standard andspecifications, before filling, the actual amount of antiserum in eachstandard reference vial will, in fact, be the same and the residualmoisture content of each vial will be the same, upon filling, as eachother vial in the same lot, because of the homogenous nature of thedried powdered bulk antiserum in the lot.

Immediately, within seconds, after filling, the vials or packages ofantiserum are sealed under sterile, dry conditions with a sealing meanswhich is absolutely dry and will not or cannot, as in past practice, hea means or source from which moisture is reintroduced into the vialedantiserum of a particular vial or package.

The use of caps, stoppers or closure members which are separately,thoroughly dried, instead of being dried simultaneously with theantiserum in the separate vials, as in the past standard practice withrespect to such standard reference reagents, ensures that there is noresidual moisture, which might otherwise, and frequently, apparently,did, remain entrapped within the cap, stopper, or closure member, whenthe antiserum content tests to the desired moisture level reenter thevial after packaging is completed to contaminate the antiserum. Thus,this invention precludes reintroduction of moisture into the antiserumafter packaging or vialing is completed.

As noted above, when the original bulk reagent is suffrciently dry, andlyophilizing is still desired, the reagent may be powdered, or at leastgranulated, before lyophilizing, if desired under all the circumstances.

Modifications, changes and improvements to the preferred and modifiedforms of the invention herein particularly disclosed and described mayoccur to those skilled in the art who come to understand the principlesand precepts thereof. Accordingly, the scope of the patent issued hereonshould not be limited to the specific forms of the invention hereinparticularly illustrated, disclosed and described but only consistentwith the advance by which the invention has promoted the art.

I claim:

1. A standard reference reagent package with a dried, homogenous,sterile, standard reference reagent therein comprising one of aplurality of such packages made from a single lot of bulk reagent by aprocess, accomplished under sterile conditions, of comminuting the bulkreagent, vacuum drying the comminuted reagent in the presence 915 adesiccant sampling the comminuted viral antiserum during the drying stepuntil the sampling indicates a uniform, homogenous moisture contentthroughout said lot of bulk comminuted viral antiserum as set bypredetermined standard and specifications, filling a plurality ofseparate, sterile, dry packages with a predetermined, standard amount ofsaid reagent from said lot in an anhydrous, inert atmosphere, andclosing each said package, immediately after filling, with a dry,moistureproof seal, while maintaining the contents of each said package,dry, sterile and uniform in accordance with said predetermined standardand specifications.

2. The package according to claim 1 in which said seal is a closuremember which is separately dried and sterilized before application tosaid package.

3. The package according to claim 2 in which said closure member ispilferproof.

4. The package according to claim 1 in which all samples of said driedlot of bulk reagent are tested for moisture content within saidpredetermined standard and specifications and within 0.2% of each other.

5. The package according to claim 1 in which said lot of bulk reagent islyophilized to a predetermined moisture content before vacuum drying.

6. The package according to claim 6 in which said lot of bulk reagent iscomminuted before lyophilizing is completed.

7. The package according to claim 1 in which said lot of bulk reagent isvacuum dried in the presence of molecular sieve.

8. The package according to claim 1 in which said lot of bulk reagent isvacuum dried in a plurality of separate containers in a single chamber.

9. The package according to claim 1 in which said bulk reagent is viralantiserum.

References Cited UNITED STATES PATENTS 2,616,604 11/1952 Folsom 53-223,035,886 5/1962 Hickey 53-22};

TRAVIS McGEl-IEE, Primary Examiner

